Tecan Journal
Life Sciences ❙ Clinical Diagnostics ❙ Forensics ❙ OEM ❙ Business and Technical News
Edition 1 / 2009
Stepping up the pace
for crop breeding projects
pages 22-23
The power is in your hands
with Freedom EVOware®
page 5
Efficient drug research
and development with the
Freedom EVO® 75
pages 18 - 19
Getting under the skin
of stem cells
pages 24 - 25
2
CEO WELCOME TECAN JOURNAL 1/2009
Welcome to an exciting New Year...
Welcome
...the Association for Laboratory Automation
(ALA) exhibition is, as ever, a great way to
welcome in the New Year. This venue is the
perfect opportunity for us to show you how
Tecan products continue to provide the
innovation, value and flexibility demanded by
your application requirements. Our newest
products include the flexible MultiChannel
Arm™ 384, improved labware logistics with
nested disposable tips, new automated
solutions for Tecan’s flagship microtiter plate
reader Infinite® M1000, and the new Cavro®
Omni Robot and Cavro® Centris Pump for
OEM customers. We are also introducing
advances in software innovation, with
easy and simple Instant Pipetting™ and
exciting Common Notification System (CNS)
Technology enabling remote monitoring
using your mobile devices, webpage, or
Windows Vista® gadgets.
Vista is a registered trademark of Microsoft Corporation.
As well as telling you more about some of
these products, this latest edition of the
Tecan Journal shows how more and more
of our customers’ challenges are being
addressed and solved by Tecan technologies.
As ever, we invite you to talk to Tecan and
see how you too can bring efficiency and
effectiveness to your laboratory.
I would like to take this opportunity to
sincerely thank you for the continued
interest, support and trust you are placing
in Tecan, our products, services and, most
importantly, in our people. We will do
everything to meet your expectations in
the coming year 2009 and jointly strive for
sustained mutual success.
We hope you enjoy this issue and, if you’re at
ALA, come and see us at booth number 205.
Thomas Bachmann, CEO
CONTENTS TECAN JOURNAL 1/2009
2
pg 7
The launch of the
regional headquarters
reflects the Company’s
significant growth and
strong commitment to
the market in China
pg 9
CEO welcome
4Infinite® M1000 and Infinite® F500
enjoy series of drug discovery assay
validations
5The power is in your hands with
Freedom EVOware®
5Tecan launches new range of
disposable tips for MultiChannel
Arm™ 96
6Tecan and GE Healthcare collaborate
for high throughput process
development
7China and Asia Pacific operations
expanded
8Investing in success
8Consolidating our organization for
customers
9Service laboratory benefits from
Tecan’s support
10-11Accuracy to the nth degree
12-13Meeting the demands of everchanging liquid handling workflows
pg 24-25
New microarray-based tool
is developed for identifying previously
difficult to diagnose primary tumors
in cancer patients
Contents
pg 18-19
14-15Semi-automated process halves
investigation time for protein-protein
interactions
16-17How stressed are you?
18-19Efficient drug research and
development with the Freedom
EVO® 75
20-21MicroRNA expression profiling of
carcinomas of unknown origin
22-23Automation accelerates crop
improvement
24-25Getting under the skin of stem cells
26-27Automated processing for pioneering
translational medicine project
28-29Signaling success
30-31Collaboration paves the way for
flexibility in diagnostic laboratories
31Get your views across with
T@lk to Tecan
32 Events 2009
3
4
PRODUCT NEWS TECAN JOURNAL 1/2009
Infinite® M1000 and Infinite® F500
enjoy series of drug discovery assay
validations
Tecan is a global leader in laboratory automation and
detection, and its high-end Infinite multimode readers enjoy
an enviable market position in terms of assay validation,
for both premium Quad4 Monochromators™- and
filter-based instruments.
Tecan is collaborating with providers of
life science and drug discovery kits and
reagents to validate its top-class multi-mode
readers, the high-end, premium Quad4
Monochromators-based Infinite M1000, and
Tecan’s fast and most sensitive filter-based
detection platform yet, the Infinite F500,
with a wide range of assay systems.
So far these include: LanthaScreen™,
GeneBLAzer®, Adapta™ Universal Kinase,
Z’-LYTE®, Omnia® PolarScreen™ and
Predictor™. Both instruments are granted
LanthaScreen Certified Plus status by
Invitrogen Corporation.
LanthaScreen™ Certified+
The Infinite F500
Transcreener® Far Red FP - the Infinite
M1000 and Infinite F500 are both
Transcreener Far Red FP validated by
BellBrook Labs, LLC.
HTRF® - both instruments have successfully
passed the HTRF compatible validation
program managed by Cisbio bioassays.
The Infinite M1000 is HTRF compatible in
white plates.
DLReady™ - both instruments have met
Promega®’s acceptance criteria for being
certified as DLReady in the corresponding
validation program managed by Promega.
This widespread validation reflects the
high performance criteria of the Infinite
systems across a wide range of detection
modules and options, and provides choice
for the customers whether to focus the work
based on monochromator or filter-driven
multimode systems.
To find out more on Tecan’s Infinite multimode
readers, visit www.tecan.com/infinitem1000.
The Infinite M1000
LanthaScreen, Adapta, PolarScreen and Predictor are
trademarks of Invitrogen Corporation. GeneBLAzer,
Omnia and Z’-LYTE are registered trademarks of
Invitrogen Corporation.
Transcreener is a registered trademark of BellBrook
Labs, LLC, USA.
DLReady is a trademark and Promega is a registered
trademark of Promega Corporation, Madison, USA.
HTRF is a registered trademark of Cisbio international,
France.
PRODUCT NEWS TECAN JOURNAL 1/2009
The power is in your hands
with Freedom EVOware®
Tecan software continues to bring reliability
and process security to users of automation.
Freedom EVOware 2.2 software makes it easier
than ever to control pipetting and scheduling
operations of Freedom EVO® liquid handling
platforms. Based on the same intuitive
graphical user interface, version 2.2 of this
software offers greater flexibility and ensures
full compatibility with the latest hardware
modules – all without compromising on
operating speed and process security.
With Instant Pipetting™, Tecan introduces
a completely new way to control pipetting
robots, without the user needing any
previous knowledge of programming.
Instant Pipetting uses a touchscreen to
enable users with any skill level to define
applications and directly control pipetting
with a liquid handling robot in real time.
This innovative feature allows the user to
perform common pipetting operations, like
sample distribution, reagent addition and
serial dilution, in a very easy and interactive
way. All steps can be repeated to perform
multiple iterations, or saved and recalled as a
Software innovation
Program
Define your pipetting
step easy and simple
Instant
Pipetting
RunControl
Recall your pipetting
easily via iPhone
Phone
RunControl
Monitor
Monitor your
instrument while
performing your
pipetting step
CNS on iPhone
or
CNS gadget
on desktop
Monitor your liquid
channel and plates
while performing your
pipetting step
iPhone
monitoring
applications
script in Freedom EVOware or mobile phone
applications, for increased ease and speed.
Once your assay is running, it is now easier
than ever for you to stay connected to your
application with the Common Notification
System (CNS). This technology enables direct
remote monitoring using your mobile phone
device, your office desktop through a Windows
Vista® gadget, or via any networked computer
with a web browser. The status of the system
is immediately visible and if user interaction is
required, it will be brought to your attention.
To find out more on Tecan’s Freedom EVOware,
visit www.tecan.com/freedomevoware.
Vista is a registered trademark of Microsoft Corporation.
Increased throughput and flexibility with
Tecan quality MultiChannel ArmTM 96
nested disposable tips
Tecan’s new disposable tip products for use
with the MultiChannel Arm 96 option for
Freedom EVO® automated liquid handling
workstations offer higher throughput
through improved logistics possibilities.
Tips without filters can be nested in passive
stacks of up to 800 tips per SBS position on
the deck, or externally in Te-Stack™ modules
to provide a capacity of 3,840 tips per Te-Stack
column, maximizing walk-away time and
improving throughput. The nested design also
offers environmental and logistical benefits,
including reduced storage space requirements
by 60 % compared to non-nested tip formats,
lower transport costs and less wastage by
using less packaging materials.
The fully SBS-compatible racks have been
designed for increased throughput and
reliability by optimizing the flexibility of
the workstation deck space. Systems fitted
with Te-Stack modules can now use the
same SBS-format columns for tip racks and
microplates, and platforms set up for nonSBS disposable tips can easily be converted.
All MCA 96™ disposable tips are produced
in a cleanroom and are certified to be
free of DNA, DNAse/RNAse, pyrogens and
endotoxins. Non-nested disposable tips are
also available in a sterile format.
To find out more on Tecan’s disposable tips,
visit www.tecan.com/consumables.
Disposable tips
stored externally in
a Te-Stack module
Disposable
tips nested in
stacks on the
workstation deck
5
6
PRODUCT NEWS TECAN JOURNAL 1/2009
Tecan and GE Healthcare
collaborate for high throughput
process development
In drug development, screening for optimal
conditions can be a tedious and timeconsuming process that can often become
a bottleneck. Time-to-clinic and time-tomarket are key factors for the success of any
drug development project, making efficient
process development a crucial component of
the overall drug development workflow.
GE Healthcare’s PreDictor 96-well plates on a
Freedom EVO workstation
With recognized expertise in their respective
fields of laboratory automation and protein
purification, Tecan and GE Healthcare are
working together to bring increased quality
and consistency to high throughput process
development. The collaboration between
the two leading suppliers focuses on
screening of protein purification conditions,
using the winning combination of Tecan’s
Freedom EVO® automated workstation with
GE Healthcare’s PreDictor™ 96-well plates
pre-filled with GE Healthcare BioProcess™
chromatography media.
The PreDictor 96-well plates have been
developed to evaluate conditions for
binding, wash and elution in a format that
allows miniaturized, parallel processing,
with the added benefits of minimal
cross-contamination and reduced sample
consumption. The good correlation of data
with those generated by chromatography
columns make the plates an excellent
tool for the initial screening of process
conditions.
PreDictor plates can be operated with
both centrifugation and vacuum protocols,
manually or in automated workflows. The
throughput, precision and robustness of the
Freedom EVO platform ensures reproducible
results from GE Healthcare’s 96-well
process development technology, providing
efficient parallel screening performed in a
multivariate fashion. Automated workflows
also offer hands-off operation and eliminate
human error, resulting in uncompromising
experimental reproducibility and consistency
among replicates. The combination of the
technologies substantially shortens time
spent on process development and reduces
overall time-to-clinic, while allowing the
investigation of a much larger experimental
space, resulting in greater understanding
of the process. The unique solution for the
optimization of process conditions provides
process developers with quality, efficiency
and reliability in downstream process
development.
To find out more on Tecan’s protein
purification solutions, visit
www.tecan.com/proteinscience.
PreDictor and BioProcess are trademarks of
GE Healthcare Companies.
CORPORATE NEWS TECAN JOURNAL 1/2009
China and Asia Pacific
operations expanded
Tecan has opened its new Asia Pacific
region headquarters, based in the heart
of Shanghai’s pharmaceutical and life
science center. The launch of the regional
headquarters reflects the Company’s
significant growth and strong commitment
to the market in China, as well as the whole
Asia Pacific region, and will improve service
and support for customers and distribution
partners alike.
The new office was opened by Thomas
Bachmann, CEO of the Tecan Group, on
the 28th October, 2008. The ceremony was
attended by key customers from around the
Asia Pacific region, major distributors, officials
and representatives from key Chinese
Provinces, the Swiss and Austrian Embassies
in China and the Tecan Group. Guests were
welcomed to the Shanghai office by the CEO
and were treated to a traditional Chinese
ceremonial inauguration of the facility,
followed by a celebratory dinner.
Thomas Bachmann thanks Peter Grünig for building up Tecan (Shanghai) Trading Co., Ltd.
Mr Bachmann said: “We are delighted to
be making this positive move into what is
almost certainly one of the fastest growing
regions in life sciences today. With a stronger
presence and new facilities, we will be able to
improve sales coverage, service and support
to all of our customers and partners in the
entire Asia Pacific region.”
Dinner with our chinese guests
Kirsti Tavast (Head of Market Unit Asia Pacific)
and Shinya Dobashi (new General Manager Tecan
Japan Co.) in front of the Asia Pacific map
7
8
CORPORATE NEWS TECAN JOURNAL 1/2009
Investing in success
Tecan is investing in focused training for
regional service managers to help them
continually improve the excellent service
support that customers have come to
expect. European service director Wilfried
Bartz has been responsible for the initiative,
and explained why he believes this training
will help provide better customer care:
“Service managers usually come from a
technical background – good engineers
with the extra talent that makes them
stand out from the crowd. The aim of this
training is to provide service managers with
a foundation of core skills to help develop
Tecan service managers and other course participants with one of the MarketingAkademie Hamburg lecturers
their regional operations. The introduction
of a course specifically for service managers
at MarketingAkademie Hamburg represents
the best training for our managers, and
offers an ideal forum to explore how
servicing challenges are handled in different
industrial environments. We specifically
chose not to have a course exclusively for
Tecan personnel in order to encourage
exchange of ideas, and focus outside of
technical issues.”
“Providing comprehensive customer
support requires more than just rapid
maintenance and servicing of hardware.
Service engineers have to be able to assess
customer needs and advise where potential
changes in practice could boost efficiency
or revenue. This ethos has to come from
the service manager and be filtered down
to the field service staff, so the benefits of
this approach need to be understood on a
management level. We also work closely
with the sales team before an order is agreed
to help provide solutions for the customer’s
needs, according to the specific application,
technical aspects and support required.
Service management is a versatile and
demanding job, and investing in our
regional managers helps them to provide
a professional and efficient service
to customers.”
Consolidating our organization
for customers
Over the past year, a small group of
representatives from the quality, finance and
other management teams within Tecan has
been working hard to improve our internal
organization and establish a more efficient,
more productive Tecan Management System.
In recent years, with the growth of the
Tecan Group, a number of different
management tools have evolved as the
size and operations of the company have
dictated. By consolidating our standard
operating processes into one integrated and
well-documented system, we have created a
strong basis from which to continue building
all organizational units and make constant
improvements within Tecan. Our new
management system will be maintained
by a dedicated department for Operational
Excellence that is directly overseen by a
member of the Executive Committee and
covers all our business organizations.
This new initiative has streamlined
decision-making processes within Tecan,
concentrating our efforts with a strong
customer focus, and we believe this new
structure will help us to respond faster, in
a more flexible way, to our customers and
the market in general. Thank you for your
continued support.
SERVICES AND CONSUMABLES TECAN JOURNAL 1/2009
Service laboratory benefits
from Tecan’s support
The Freedom EVO 100 has been integrated with a microplate heat sealer and is accessed by a robotic arm
The Cellular Screening Center (CSC) in the Institute for
Genomics and Systems Biology (IGSB) at the University of
Chicago provides a cell screening service for drug discovery
and development in several local faculties. The reliability of
Freedom EVO® workstations, supported by Tecan’s excellent
customer service, copes easily with high throughput liquid
handling for various research projects.
Sam Bettis, Technical Director at the CSC,
explained: “We are a service laboratory for
researchers here at University of Chicago,
as well as at University of Illinois at Chicago
and at Northwestern University, performing
their high throughput analysis projects such
as whole genome siRNA screening and small
molecule screening.”
Starting with a clean sheet, Sam and
his colleagues designed the laboratory
and selected the automation robotics,
including two Freedom EVO liquid handling
Sam Bettis, Technical Director, and Nicolette
Pavlovics, Research Technologist, study the
Freedom EVO 100 through its PC terminal
workstations. The first Freedom EVO 100 has
an integrated liquid handling (LiHa) arm with
eight independent tips, and is capable of
constructing rearrays of RNAi master plates as
well as dispensing into 96- and 384-well plates.
In addition, a Freedom EVO 200, equipped
with a LiHa arm with eight tips, a robotic
manipulator (RoMa) arm and a Te-MO™384,
is used for plate set-up and replication.
Sam continued: “The modularity of
Tecan platforms allowed us to upgrade
the Freedom EVO 200 by integrating a
microplate heat sealer, which now sits
to the right of the instrument and is
accessible with the RoMa arm. We are also
in the process of integrating a laboratory
information management system that
will allow us to streamline siRNA storage
and retrieval. It will automatically export
Microsoft Excel® files to allow cherry picking
with the Freedom EVO, which will be great.”
“I chose the Freedom EVO platforms because
I had worked with Tecan instruments
previously, and talking to many other people
The Freedom EVO workstations at the CSC are able
to process 384-well plates
in the field confirmed that apart from
having the best reliability and precision,
Tecan offered excellent service. Our
laboratory can’t afford any downtime, so
services like after-care support and annual
service contracts are very important.”
“We have a complex set-up, so Tecan has
helped us with some software-related
tasks such as integrating the sealer with
the Freedom EVO 200, and for the software
upgrades of the Freedom EVO 100 to ensure
that communication with the robotic arm
is maintained. Tecan always responds very
quickly and we have a fantastic service
engineer located here in Chicago. Everybody
at Tecan from sales to service has been
excellent but generally, once we had become
familiar with the instruments, there have
been few issues; it turns out that we’re much
better at breaking the robots than the robots
themselves are!” Sam concluded: “We are very
happy with the Tecan workstations, and I have
recommended them to several laboratories.”
Excel is a registered trademark of Microsoft Corporation.
9
10
LIQUID HANDLING TECAN JOURNAL 1/2009
Accuracy to the nth degree
Researchers at Quanterix in
Cambridge, Massachusetts,
USA, are relying on a Freedom
EVO® 150 liquid handling
workstation to provide reliable
and consistent pipetting in the
development of a revolutionary
new technology for single
molecule detection.
An optical fiber bundle. The higher
magnification view (inset) shows the
uniform individual reaction vessels
Quanterix Corporation in Cambridge,
Massachusetts, USA, was founded in 2007
to develop a novel platform for single
molecule detection. The revolutionary
SiMoA™ (Single Molecule Array) technology
was originally developed at Tufts University,
Massachusetts, by Quanterix scientific
founder Dr David Walt, and is based on
arrays of femtoliter-sized reaction vessels
arranged uniformly on the tip of an optical
fiber bundle. Each reaction vessel is isolated
from neighboring vessels, and when
incubated with dilute solutions, can be
used to trap single molecules according to
Poisson statistics. Fluorescent signals are
detected via a custom instrument using
automated image analysis software.
Initial applications for this innovative
technology are focused on the detection
of protein biomarkers. Quanterix has
combined single molecule detection with
the reagents used in immunoassays to
achieve a 1,000-fold increase in sensitivity
over standard ELISA. The company hopes
that enhanced sensitivity will lead to
real clinical benefits for patients. David
Hanlon, Director of Strategic Marketing and
Collaborations at Quanterix, explained: “This
technology employs very similar principles
Fluorescent image of the optical fiber
bundle. Illuminated spots represent single
protein molecules
to traditional ELISA methods, but instead of
detecting many molecules per well as with
traditional analog systems, our technology
isolates individual molecules in femtoliter
volume reactions for a digital output. This
approach offers a significant increase in
sensitivity relative to standard assays, so
we are looking into applications where
the improved detection limits will provide
real value to clinical diagnostics. There are
many clinical opportunities for improved
biomarker detection, including the detection
of proteins implicated in oncology and
cardiovascular disease, as well as markers
useful for monitoring infectious agents
and inflammatory response. One exciting
prospect is the possibility of switching
from detection of biomarkers in blood to
less invasive body fluids, where the same
marker proteins may be present at much
lower concentrations. Using the sensitivity
of the SiMoA platform, we are hoping to
characterize the expression of many of these
biomarkers in saliva or urine. We expect
that our technology could be of particular
value for screening programs, where earlier
diagnosis of a condition could potentially
have a much more favorable outcome for
the patient.”
LIQUID HANDLING TECAN JOURNAL 1/2009
Todd Campbell, Senior Automation Engineer,
continued: “We currently use the Freedom
EVO platform as a research and development
tool to optimize our assays, ensuring
consistency throughout every assay we run
from day to day. We use the automated
system for both liquid handling as well
as the manipulation of our consumable
test strips. With the assay protocols under
continuous evolution, this makes it much
easier to make small adjustments to
the system. The Freedom EVO offers the
flexibility we need to make these changes
as necessary, and the software is easy to use.
This has enabled us to develop our assay
platform more quickly, and begin testing
samples with our instrument.”
Jeffrey Randall, Principal Scientist, explained
why Quanterix opted for Tecan systems: “Our
proprietary assay and detection system is at
least 1,000 times more sensitive and much
more accurate than the traditional bulk ELISA
assays, so careful control of pipetting volumes
is important to us. The Freedom EVO platform
Todd Campbell holding an optical fiber bundle
offers very repeatable pipetting, and this
has allowed us to ramp up our throughput
without sacrificing the consistency of our
results. We looked at other automated liquid
handling systems on the market, but they did
not meet our requirements.”
In his role as senior automation engineer,
Todd has worked closely with Tecan to
optimize the Freedom EVO platform
for Quanterix’s needs. “I have been very
impressed with Tecan’s support. When we
were first setting up the system, our sales
engineer was very helpful and extremely
knowledgeable about the system. In the
past, I have often found that equipment
manufacturer’s sales personnel are not
necessarily system specialists, but that
is certainly not the case with Tecan. With
our Freedom EVO systems in heavy use
every day, it is especially important that
any issues we have are resolved quickly.
Whenever we have needed support, they
have been here within 24 hours, which is
very impressive.”
11
(l to r) David Rissin, Senior Scientist,
Jeffrey Randall, Principal Scientist, and Todd Campbell,
Senior Automation Engineer, with the Freedom EVO
SiMoA is a trademark of Quanterix Corporation.
12
APPLICATION BIOPHARMA TECAN JOURNAL 1/2009
Meeting the demands
of ever-changing liquid
handling workflows
The Bioprocess, Analytical
and Formulation Sciences
Group at Merck, West Point,
Pennsylvania, USA, has
accumulated several Tecan
instruments over the years
for the development and
automation of high
throughput biological assays,
essential in the support of
bioprocess development of
vaccines and biological products.
The Bioprocess, Analytical and Formulation
Sciences Group at the Merck research
and manufacturing facility at West Point,
Pennsylvania, develops biological methods
for a number of processes involved in the
production of therapeutic proteins and
vaccines. Over several years, the group has
built up a wealth of Tecan liquid handling
and detection systems and continues
to develop new and innovative ways of
automating its ever-changing workflow.
Kristine Little, Research Biochemist, said: “We
use our Tecan systems to develop automated
assays and provide a routine service in
testing and processing to laboratories within
the West Point site using these assays. We
also carry out activity and binding assays for
final products of the purification processes;
biochemical testing including assays for
residual DNA and concentrations of proteins
and process residuals; assays for residual
host cell proteins and binding assays such as
EC50 or IC50; and ELISA assays.”
“We have accumulated our collection of
14 Tecan systems over the past decade,
including several Freedom EVO® platforms
equipped with microplate readers and the
Power Washer 384™ microplate washer for
processing immunoassays, and some older
Genesis RSP™ liquid handling platforms, also
incorporating different add-on modules,”
Kristine continued. “The decks on all the
instruments are set up to give us maximum
flexibility so we can run a variety of assays
on any workstation. Some are grouped to
run immunoassays, whereas others run
the protein and DNA assays – the ‘mix
and read’ type of simple one- or two-step
assays that can easily be automated.
These usually involve adding dye and
buffer mixture to the sample, followed by
mixing and measurement of fluorescence
intensity, giving sensitive, selective and quick
quantification of nucleic acids and proteins.”
Amy Bowman, Research Biochemist,
continued: “The Tecan systems dedicated
to ‘mix and read’ DNA and protein assays
are set up to run five or six plates per
run unattended, and we have a similar
throughput for immunoassays of six plates
at a time. Only one analyst is needed to
Tecan’s Te-Chrom™ module, developed in close collaboration with Atoll GmbH, allows efficient automated processing of Atoll’s MediaScout RoboColumns
on the Freedom EVO platform. When combined with the Te-Stack™ module, the Te-Chrom is able to collect fractions of volumes as low as 25 μl.
APPLICATION BIOPHARMA TECAN JOURNAL 1/2009
(l to r) Kristine Little and Amy Bowman with their
back-to-back Tecan system
set up the Tecan instrument and then,
depending on the assays, the run time
varies from two to 14 hours. The automated
systems are in full-time use and the six
analysts responsible for running the processmonitoring laboratories will often re-set
more than one instrument on the same day.”
“Throughput and flexibility are the main
reasons we continue to choose Tecan and
why we have recently added even more
systems to our collection. Our most recent
acquisition is a new back-to-back system,
built and customized for us by the Tecan
Integration Group (TIG) in North Carolina,
and consisting of a Freedom EVO 200
platform on the front and a Freedom EVO 150
platform at the back. Together these platforms
will run various ELISA assays as a carouselbased, high throughput system, handling as
many as 1,440 analyses in 20 96-well plates,
unattended over the course of 24 hours.”
“For us, flexibility in programming is just as
important as in the hardware; samples come
to us in a diverse range of types and formats,
and we need to be able to handle different
numbers and types of samples from one
run to the next,” Amy added. To achieve
this we have written and developed some
Visual Basic® software in-house so we can
create customized worklists for our sample
runs. It is also important that Tecan offers
the choice of so many different devices
compatible with its liquid handling systems.
We use several modules incorporated into
our Genesis RSP and Freedom EVO platforms,
including numerous Te-MO™ multichannel
pipetting arms, four MCA™ 96 multichannel
arms and one MCA™ 384. The multichannel
arms allow us to increase throughput and
will enable us to make use of microfluidic
devices – for performing small-scale ondeck microscale processes – in our future
development of purification methods.
We recently purchased the first of these
devices – a MediaScout® 96-array
RoboColumn high throughput purification
column system from Atoll – and have
successfully integrated this into one of our
Freedom EVO platforms. This will carry out
chromatography analysis, specifically high
throughput screening of chromatographic
resins, which is the next step in our
expansion plans for the whole set-up.”
Visual Basic is a registered trademark of Microsoft
Corporation.
MediaScout is a registered trademark of Atoll GmbH.
13
14
APPLICATION BIOPHARMA TECAN JOURNAL 1/2009
Semi-automated process
halves investigation time for
protein-protein interactions
The Department of Molecular
Pharmacology at the Marseille
Cancer Research Center,
France, has developed a semiautomated yeast two-hybrid
system with the Te-MO™ 96
Multichannel Pipetting Option
from Tecan, halving the time
taken to obtain reproducible
results from six to three days.
The state-of-the-art Marseille Cancer
Research Center has developed a strong
interest in the molecular biology of cancer.
The Center’s molecular pharmacology
laboratory is involved in ongoing
investigations into potential therapeutic
targets, using yeast two-hybrid (Y2H)
screening techniques for the discovery
of protein biomarkers. The Y2H system is
a well-established method for screening
of protein-protein interactions (PPIs),
relying on PPI-dependent activation of a
reporter gene for identification of positive
clones. Historically, false positive results
have been the major drawback of Y2H
techniques, requiring multiple replicates
and protracted cleaning procedures to
ensure reproducibility. In order to streamline
this process, the molecular pharmacology
laboratory has developed a semi-automated
protocol for Y2H screening.
Patrick Lécine with the
Freedom EVO
Comparison of the phenotypic assay results.
Phenotypic assays were performed either with the replica-cleaning (upper panel) or the
liquid dilution protocol (lower panel) to compare their efficiency in detecting positive
interactions using the four phenotypic assays
Frederique Lembo inserts
a plate for the Te-MO 96
Patrick Lécine is responsible for all yeastbased screening at the Center, and
explained why they chose to develop a
semi-automated protocol: “Manual replica
plating for Y2H analysis requires plating
out an excess of yeast, far more than is
necessary for phenotype determination,
in order to guarantee reproducibility. To
eliminate false positives, several cleaning
steps with a replica block are needed to
remove the excess yeast. These are timeconsuming, repetitive, tedious and prone
to experimenter-dependent factors, such
as the amount of pressure applied to the
replica block and the number of replica
cleaning steps required. We have automated
the replica plating protocol using a Te-MO 96
Multichannel Pipetting Option from Tecan,
which improves colony definition and
distribution for unambiguous phenotype
analysis, and allows us to use a simplified
dilution protocol and less yeast culture per
colony. Cross-contamination is avoided and
reproducibility is improved significantly;
APPLICATION BIOPHARMA TECAN JOURNAL 1/2009
the multiple replica cleaning steps are no
longer required but we get the same results
and the Y2H phenotyping assays are
much faster.”
Patrick continued: “Yeast cultures are created
with plasmids encoding a transcription
factor (Gal4) fused to bait sequences, and
a number of reporter genes requiring Gal4
activation. These are transformed with
different sequences from cDNA libraries,
then grown in modified synthetic complete
(SC) liquid medium deficient in leucine,
tryptophan and histidine. Positive clones
are selected and re-plated in 96-well format
on solid SC medium for four to five days to
confirm the phenotype, before transfer to
liquid SC medium deficient in leucine and
tryptophan. After three days growth cultures
reach stationary phase, and concentration
is determined across all wells based on OD600
absorbance measurements. From this point the
procedure is automated using the Te-MO 96.
Cultures are gently
re-suspended and diluted, before plating
cultures onto selective SC media for
identification of positive clones of one of
three phenotypes (His+, Ura+ and 5-FAOr).
In addition, 5 μl of undiluted culture is plated
onto a nitrocellulose filter (YPD) plate for
the β-galactosidase activity assay. Starting
with one 96-well donor microplate, the resuspension, dilution and plating out of yeast
cultures takes only six minutes. It has been
relatively simple to automate the pipetting
steps on the Te-MO 96, thanks to the easyto-use Gemini™ software, and our semiautomated protocol can be used for any Y2H
screening application by varying the yeast
starter culture or the phenotyping media in
the plates.”
“The reproducible pipetting of the TeMO 96 has allowed us to adopt this new
dilution-based replica plating process,”
Patrick concluded, “making automation
possible by eliminating the need for replica
block cleaning of plates, halving the time
required to identify PPIs and minimizing the
volumes of liquid and solid media required.
However, the current protocol is limited
by the incubation time on solid media
required to achieve equal yeast uptake
with each tip of the manual multichannel
pipette. In the future, we intend to put the
Te-MO unit together with our Freedom
EVO® 150 workstation in order to improve
the reproducibility of pipetting for smaller
colonies, allowing us to reduce incubation
times. This will also allow us to automate
OD600 measurements for liquid colonies,
allowing absorbance to be measured and
dilutions to be normalized after only 24
to 36 hours of incubation. Together, these
improvements should both reduce the
total time required for phenotype analysis
and give rise to a fully automated process.
The team at Tecan has been good to work
with, and someone is always available for
help and advice, so we hope to achieve full
automation soon.”
Phenotypic assays obtained with liquid dilution of the controls. Phenotypic assays
were performed on SC-WHL + 25 mM 3-AT and SC-WUL plates using the seven
controls and a serial dilution starting from a OD600=2 to 0.001.
15
16
LIQUID HANDLING TECAN JOURNAL 1/2009
How stressed are you?
Psychologists at Anglia Ruskin
University, UK, are using a
Tecan liquid handling
platform to automate
ELISA-based analysis of
cortisol and IgA levels in
their investigations into the
relationship between different
types of stress and the
immune response.
Researchers in the Department of
Psychology at Anglia Ruskin University, UK,
are exploring the relationship between
stress and responses of the human
body, particularly the immune system,
using specific examples to look at the
differences between chronic and acute
stress. Dr Matt Bristow, Senior Lecturer
in Biological Psychology, explained: “We
are concentrating on various groups of
individuals, effectively trying to model
stress in a controlled environment and
looking closely at the immune responses of
those individuals. For example, one of the
most recent projects was a collaborative
study into the experiences of carers –
usually husbands, wives or partners – of
patients with frontal temple dementia, a
type of dementia similar to Alzheimer’s
disease but with an earlier average age
of onset of 55 years. Our initial study
found that carers who reported longterm chronic stress tended to have higher
levels of immunoglobulin A (IgA) rather
than the lower levels we expected. These
surprising findings may indicate that the
immune systems of carers are reacting
to the challenges of their day with large
increases, or alternatively that IgA levels
are not suppressed by chronic stress as
was previously thought. This on-going
research reveals the potential for salivary
biomarkers to help us understand the health
implications of real-life stressful situations.”
“At the other extreme, we followed a
number of volunteers through their first
tandem parachute jump, and mapped the
response of the immune system to this kind
of acute stress. There were progressively
huge increases in cortisol in their saliva at
each step, starting with the day before the
event, arriving at the parachuting centre,
going onto the aircraft and lastly, the actual
jump from the aircraft at 10,000 feet. A
large, transient increase in their immune
responses, measured by levels of IgA, the
most abundant antibody type in saliva,
follows the rise in cortisol. This is consistent
with previous literature; in response to a
brief challenging event such as acute stress,
the immune system reacts by releasing
natural killer cells, IgA and proteins. The
levels of these drop off extremely quickly,
with IgA levels returning to normal within
40 minutes, but cortisol is still high hours
after the jump.”
LIQUID HANDLING TECAN JOURNAL 1/2009
“In another study, we discovered that, in a
10-day period, stressed undergraduates tend
to have very volatile IgA levels, whereas
their non-stressed counterparts have
steady IgA levels, giving a flat profile. It is
clear from this and other examples, that
studying an immune profile over several
days gives a much better indicator of stress
levels compared to taking just a single
measurement. We need to have a thorough
understanding of variability in immune
responses and therefore collect saliva
samples several times a day for up to two
weeks, creating thousands of samples that
would be impossible to handle manually.”
Matt continued: “We have used a Tecan
platform to automate our ELISAs since
2006 and our daily throughput has
increased considerably, from 70 to 100
samples manually to over 600 on the Tecan
system. We have purposefully restricted
ourselves to handling only saliva samples,
as they are so much easier to collect than
blood, and indeed can be collected by
participants themselves, with far fewer
worries about storage and health and
safety. There are also now so many more
biomarkers of interest detectable in saliva,
including antibodies, DHEA, testosterone
and C-reactive protein, a marker in heart
disease. The system includes a PosID™
scanner for barcode recognition and AIS
LIMS software for tracking, integration
with databases and data printing so, once
the samples are loaded and the assays
start, we can just leave it running. We are
required by Human Tissue Act legalization
to track all human tissue samples through
the laboratory and barcoding gives us
complete certainty that no sample mix-up or
mislabeling has occurred. In all, the system
eliminates human errors, schedules assays
dynamically and very effectively to increase
throughput, and gives us much more
reliable data than before, in a far shorter
time. I am very pleased that we managed
to get a fully automated system that has
delivered considerably more than our initial
expectations.”
“The Tecan platform has given us extra
capabilities and we are not yet running it
at maximum capacity,” Matt concluded.
“We have already built up several academic
collaborations with other research teams
and units using the equipment. This
consultancy service brings in more capital
for the university and our department,
allowing us to continue our research
without worrying too much about funding,
and the high capacity of the system means
we still have room to deal with any studies
that come our way.”
17
18
APPLICATION BIOPHARMA TECAN JOURNAL 1/2009
Efficient drug research and
development with the
Freedom EVO® 75
Toa Eiyo in Japan is a
pharmaceutical company
specializing in the research
and development of
pharmaceutically unique
medicinal products for
cardiovascular diseases such
as angina or heart attacks.
The Company’s
pharmacokinetic testing
laboratory in Fukushima has
chosen the Freedom EVO 75
equipped with an 8 Plus 1
Access™ Liquid Handling Arm
to achieve greater efficiency
by automating its sample
preparation steps.
The development of new drugs at
Toa Eiyo starts with the synthesis and
evaluation of various novel low molecular
weight compounds at its Tokyo Research
Laboratories. The compounds that
demonstrate potency are selected for
pharmacokinetic testing at Toa Eiyo’s
Fukushima Research Laboratories. Mr
Yoshiaki Watanabe, chief scientist, explained:
“Preliminary pharmacokinetic studies
using cells of human origin or artificial
membranes are performed at our Tokyo
Research Laboratories, and promising
compounds are transferred to our section for
further pharmacokinetic, toxicity and safety
testing. Our section has around 20 members,
of whom six are involved in pharmacokinetic
studies, involving measurement of drug
concentration in blood plasma from animal
testing and clinical trials. In addition, Toa
Eiyo’s Pharmaceutical Technology Center
is making special efforts to develop drug
formulations that are pharmaceutically
unique, and pharmacokinetic profiles of
these new formulations are also evaluated
in our section. With this recent increase
in the number of samples we analyze, we
wanted to automate the sample preparation
steps to achieve greater efficiency. Tecan’s
liquid handling devices greatly attracted
our interest at the 8th International ISSX
Meeting in October 2007, and we selected
the Freedom EVO 75 with integrated 8 Plus 1
Access Liquid Handling Arm, which offered
the management of pipetting precision with
its liquid level detection system.”
They use the Tecan instruments for
automated preparation of blood plasma
samples, which involves solid phase
extraction (SPE) or deproteinization in 96well filter plates, using customized programs
for each step. As pipetting precision is
critical, an integrated balance is used before
each run to confirm the pipetting precision
of the first channel, which is used for sample
transfers, while all eight channels are used
for other preparation tasks. Mr Watanabe
Toa Eiyo’s Pharmaceutical Technology Center
develops pharmaceutically unique formulations
for the treatment of heart disease.
Photo provided courtesy of Toa Eiyo
Toa Eiyo’s pharmacokinetic team (l to r, back row): Mr Yoshiaki Watanabe, Mr Kazuhiro Taniyama,
Mr Takashi Sato (l to r, front row) Ms Sakiko Suzuki, Ms Marie Tanaka, Mr Toshihisa Kodama
!00,)#!4)/.ß")/0(!2-!ßßßß4%#!.ß*/52.!,ß
ISßCONSIDERINGßTHISßSTEPßASßAßUSEFULßTESTßFORß
COMPLIANCEßWITHß'OODß,ABORATORYß0RACTICEß
STANDARDSß4HEß,IQUIDßLEVELßDETECTIONßSYSTEMß
ONßTHEß½RSTßCHANNELßHELPSßTOßPREVENTß
SAMPLINGßERRORßANDßMINIMIZEßWASTAGEß
OFßVALUABLEßSAMPLESßFURTHERßENHANCINGß
PIPETTINGßRELIABILITYß*USTßBYßSETTINGßBLOODß
PLASMAßSAMPLESßINßTHEß&REEDOMß%6/ßßTHEß
PREPARATIONßISßCOMPLETEDßAUTOMATICALLYßAFTERß
WHICHßTHEßSAMPLESßAREßSIMPLYßTRANSFERREDßTOß
THEß,#-3-3ßFORßANALYSIS
±4HEßAUTOMATEDßPREPARATIONßOFßONEßPLATEß
WITHßßPLASMAßSAMPLESßTAKESßANßHOURßANDß
AßHALFßTOßTWOßHOURSßWHEREASßBYßMANUALß
PIPETTINGßITßUSEDßTOßTAKEßATßLEASTßHALFßAßDAY²ß
-Rß7ATANABEßELABORATEDß±4HEßPREPARATIONß
STAGEßISßNOßLONGERßAßBOTTLENECKßBECAUSEßTHEß
AUTOMATEDßPROCESSßISßFASTERßTHANßTHEß,#-3
-3ßANALYSISß4HEß&REEDOMß%6/ßßCOULDß
PROCESSßFOURßORß½VEßPLATESßINßAßDAYßBUTßWITHß
THEßCURRENTßSETUPßWEßPROCESSßTWOßORßTHREEß
PLATESßINßAßDAYßANDßANALYZEßTHEßSAMPLESß
OVERNIGHTßONßMULTIPLEß,#-3-3ßSYSTEMSßFORß
THEßRESULTSßTOßBEßCOLLECTEDßTHEßNEXTßMORNING²
!NOTHERßOBJECTIVEßOFßINTRODUCINGßTHEß
AUTOMATEDßALIQUOTINGßDEVICEßWASßTOßß
PREVENTßHUMANßERRORß-Rß7ATANABEß
CONTINUEDß±5NTILßRECENTLYßWEßHAVEßBEENß
HANDLINGßAßLARGEßNUMBERßOFßSAMPLESß
MANUALLYßPAYINGßAßGREATßDEALßOFßATTENTIONß
TOßMINIMIZEßMISTAKESßBUTßBYßAUTOMATIONß
THATßKINDßOFßHUMANßERRORßCANßNOWßBEß
PREVENTEDß!UTOMATIONßHASßALSOßREMOVEDß
THEßPSYCHOLOGICALßBURDENßONßTHEßRESEARCHERSß
WITHßAßSMALLßNUMBERßOFßPEOPLEßPARTICIPATINGß
INßAßWIDEßVARIETYßOFßTASKSßAUTOMATIONßCANß
CREATEßMOREßSPACEßINßTHEßSCIENTISTS´ßTIMEßANDß
WEßANTICIPATEßTHATßWEßWILLßBEßBETTERßABLEßTOß
PLANßOURßRESEARCHßTHEMESß7EßHAVEßALREADYß
SEENßSAVINGSßINßTIMEßANDßTHEßAMOUNTßOFß
PRECIOUSßSAMPLESßUSED²
±4HEßCHARACTERISTICSßOFßTHEß&REEDOMß%6/ß
MAKEßITßEASYßTOßUSEßFORßMANYßRESEARCHERSß
%VENßWITHOUTßSPECIALISTßKNOWLEDGEßITßISßß
VERYßEASYßTOßWRITEßAßSCRIPTßINß&REEDOMß
%6/WARE‡ßANDßITßISßPOSSIBLEßTOßADAPTß
EXISTINGßPROGRAMSßFORßOTHERßMANIPULATIONSß
WITHßJUSTßAßFEWßCHANGES²
-Rß7ATANABEßISßPLANNINGßTOßUSEßTHEßß
&REEDOMß%6/ßßINßOTHERßPROJECTSß±7EßWOULDß
ALSOßLIKEßTOßAPPLYßOURßWORK¾OWßTOßOTHERß
PRECLINICALßTESTSßTOßEVALUATEßNUMEROUSß
EVALUATIONßCOMPOUNDSßEF½CIENTLYßLIKEß
SOLUBILITYßSCREENINGßMETABOLICßSTABILITYß
TESTINGßETCß4HESEßAREßCURRENTLYßBEINGßCARRIEDß
OUTßMANUALLYßBUTßWEßWOULDßLIKEßTOßAUTOMATEß
ASßMANYßOFßTHEßROUTINEßTASKSßASßPOSSIBLE²
±&ORßTHEßEF½CIENTßPROGRESSßOFßRESEARCHß
PROJECTSßITßBECOMESßCRITICALßTHATßEACHß
RESEARCHßWORKERßCANßMAKEßDECISIONSßBASEDß
ONßTHEßDATAßOBTAINEDßANDßTHEßINSTALLATIONßOFß
ANßAUTOMATEDßPIPETTINGßDEVICEßTHATßCANßBEß
APPLIEDßTOßPRECLINICALßANDßCLINICALßTESTSßHASß
BEENßVERYßWELCOMEß)TßHASßALREADYßHELPEDß
TOßREDUCEßTHEßBURDENßOFßROUTINEßMANUALß
TASKSßANDßWEßANTICIPATEßAßFARREACHINGßEFFECTß
OFßITSßUSEßINßOURßFUTUREßRESEARCHßTHEMES²ß
CONCLUDEDß-Rß7ATANABE
&IRSTßESTABLISHEDßINßß4OAß%IYOßHASß
ALWAYSßBEENßAßRESEARCHßANDßDEVELOPMENTß
ORIENTEDßCORPORATIONßUTILIZINGßTHEßVITAMINß
"ßCONTAINEDßINßSILKWORMßPUPAEßTOßBECOMEß
THEßTOPßMANUFACTURERßOFßVITAMINß"ß
FORMULATIONSßINß*APANß)NßTHEßSß4OAß
%IYOßSUCCESSFULLYßDEVELOPEDßHEARTßDISEASEß
TREATMENTSßWHICHßWEREßIMMEDIATELYß
PRAISEDßBYßMANYßINßTHEßMEDICALß½ELDßASß
UNIQUEßANDßEFFECTIVEßPRODUCTSß4ODAYß
4OAß%IYOßSPECIALIZESßINßTHEßDEVELOPMENTß
OFßDRUGSßFORßUSEßINßTHEßTREATMENTßOFß
CIRCULATORYßANDßCARDIOVASCULARßCONDITIONSß
SUCHßASßISCHEMICßHEARTßDISEASEßINCLUDINGß
ANGINAßANDßMYOCARDIALßINFARCTIONßANDßISß
DEDICATEDßTOßSUPPORTINGßANDßENHANCINGß
THEßQUALITYßOFßLIFEßOFßPATIENTS
-ANYßNOWßLOOKßTOßTHEßCOMPANYßTOß
DELIVERßHOPEßTOßPATIENTSßWITHßHEARTß
DISEASEß4OAß%IYOßISßCOMMITTEDßTOßPURSUEß
EVERßHIGHERßLEVELSßOFßORIGINALßRESEARCHß
ANDßDEVELOPMENTßATßITSßFACILITIESß
AIMINGßTOßBECOMEßAßLEADINGßSPECIALIZEDß
PHARMACEUTICALßCOMPANYßWITHßAßTRULYß
VALUEDßROLEßINßTHEß½ELDßOFßCIRCULATORYßANDß
CARDIOVASCULARßTREATMENTß4HEßDEMANDßFORß
BETTERßANDßSAFERßMEDICINESßTOßTREATßADULT
RELATEDßDISEASESßWILLßCONTINUEßTOßINCREASEß
ANDß4OAß%IYOßISßRESPONDINGßTOßTHEßNEEDSßOFß
THEßAPPROACHINGß³ERAßOFßTHEßELDERLY´ßASßITSß
MOSTßIMPORTANTßSOCIALßOBJECTIVE
20
MICROARRAY PRODUCTS TECAN JOURNAL 1/2009
MicroRNA expression profiling of
carcinomas of unknown origin
Loading samples into the HS 4800 Pro hybridization station in the service department at Exiqon
Around 5 % of all newly diagnosed metastatic cancers are
carcinoma of unknown primary (CUP) origin, where the site of
the primary tumor cannot be determined, despite the use of
advanced immunohistochemical or radiological techniques.
Because effective cancer treatment depends on early
identification of the primary tumor, CUP patients have poor
prognoses, with a median survival of 3-6 months and a oneyear survival rate of less than 25 %.
Exiqon has developed a new molecular tool
for the diagnosis of CUP, using microarraybased microRNA profiling data. MicroRNAs
are good candidates for such a tool as their
expression signatures can be used to classify
specific cancers. Simultaneous detection
of all microRNAs on an array presents a
number of challenges. The short length
(21-23 nt) of microRNAs leaves little room for
probe optimization, and many microRNAs
are very similar in sequence, varying by only
one or two bases. Also, microRNAs possess
huge variation in base composition, which
results in a large TM range of microRNADNA probe dimers (Figure 1, gray bars). To
overcome these challenges, the capture
probes used in the miRCURY™ LNA™
microRNA Arrays incorporate Exiqon’s
high affinity Locked Nucleic Acid (LNA)
technology. The TM-normalized probes
(Figure 1, yellow bars) of these arrays result
in unparalleled sensitivity and excellent
mismatch discrimination for all microRNAs,
making them superior to DNA probes (Figure
2) for efficient discrimination between
closely related microRNA family members
and microRNA profiling from as low as 30 ng
total RNA.
Automation is an important factor for
standardization of complex experimental
procedures, such as those involved in
hybridization experiments, to ensure
reproducible and reliable data. Tecan’s HS Pro
series of hybridization stations – available
as HS 400™ Pro, which handles up to four
slides, and a larger version, HS 4800™ Pro,
which can process up to 48 slides – enables
full automation of microarray hybridization
experiments with minimal handling of
solutions and slides. The hybridization
station applies washing buffer to the active
slide surface within a hybridization chamber
that seals the slide from the top to improve
washing efficiency. During hybridization,
the sample is subjected to the patented
agitation mechanism to guarantee uniform
incubation. The agitation, together with
hybridization station’s unique active
bubble suppression (ABS™) system, helps
to achieve maximum specificity, sensitivity
and reproducibility of results. The on-slide
nitrogen drying procedure (OSND™) results
in a low and uniform background, and the
slides can be scanned immediately without
the need for tedious drying procedures.
Exiqon developed the miRCURY LNA
microRNA Arrays using the HS 4800 Pro, and
three are in its R&D laboratories for product
development. Another seven HS 4800 Pros
are being used in the service department
and are operated to full capacity, every day,
where samples received from customers
are processed. Although Exiqon’s arrays can
be hybridized manually, the Company has
always recommended automated processing
using Tecan’s hybridization stations for the
most reliable results.
To develop a classifier tool for diagnosis
of CUP, Exiqon’s miRCURY LNA microRNA
Arrays were processed on Tecan HS 4800 Pro
hybridization station, for microRNA profiling
of normal and tumor tissues. More than 500
tumor and normal adjacent tissue samples
were collected from both fresh frozen and
formalin fixed, paraffin embedded sections
of 18 tissue types (adrenal, bladder, breast,
cervix, colon, esophagus, gall bladder, kidney,
liver, lung, ovary, pancreas, prostate, rectum,
small intestine, stomach, testis and uterus),
representing the most common tissues of
origin for CUP. Total RNA from tissue was
analyzed for microRNA expression on the
MICROARRAY PRODUCTS TECAN JOURNAL 1/2009
Figure 1. TM-normalized LNA capture probes. DNA
capture probes (gray bars) for human microRNAs
have a TM range of more than 30 °C and an average
TM of 64 °C (StDev = 5 °C). The LNA capture probes
(yellow bars) have a TM range of only 10.0 °C and
an average TM of 71.5 °C (StDev = 1.6 °C).
Figure 2. LNA-based arrays are superior to DNAbased arrays at detecting microRNA. Upper
panel: Exiqon’s LNA-based array. Lower panel: A
DNA-based competitor array. Gray bars indicate
the percentage of capture probes capable of
detecting 50 amol of synthetic microRNA within
the given range of GC content. Yellow bars indicate
the percentage of capture probes not detected at
50 amol. It is clear that DNA-based arrays show
decreasing efficiency with decreasing GC content.
miRCURY LNA microRNA Array using the
HS 4800 Pro hybridization station, and a
unique microRNA profile for each of the
18 tissue types was generated (the full
protocol for hybridization and washing can
be downloaded at www.tecan.com or at
www.exiqon.com). A microRNA expression
database was established, and a detailed
analysis of these results revealed that a
cancer classifier based on the expression
levels of seven microRNAs is sufficient to
distinguish between tumors from the 18
tissue types.
The classifier tool was subsequently used
to identify the tumor type of a patient
suffering from CUP by analyzing a biopsy
from a lymph node metastasis. A biopsy
from a lymph node metastasis from a
patient suffering from CUP was profiled
on the miRCURY LNA microRNA Array. The
microRNA expression level of the metastasis
was compared to the average expression
level across the 18 tissues and, based on
a simple cluster analysis, the lymph node
metastasis microRNA profile was shown to
belong to the colorectal microRNA cluster
(Figure 3). A primary colon tumor was later
found in this patient, which highlighted
the potential of this method as a tool for
identifying primary tumors in CUP patients.
Therefore, in this case, one would – based
on the microRNA profile alone – be able to
direct the treatment against a colon cancer
without having to perform a full body PET
scan to search for the primary tumor. A
colonoscopy would have been sufficient to
verify this result.
Taken together, the results indicated that
the miRCURY LNA Array used in conjunction
with the HS 4800 Pro hybridization station is
a very powerful combination for high quality
microRNA expression studies. It permits
highly specific probe binding by yielding
maximum sensitivity and a high inter- and
intra-slide reproducibility by circumventing
the handling drawbacks and the decrease
in data quality associated with manual
hybridization experiments.
LNA and miRCURY are trademarks of Exiqon A/S.
Figure 3. The workflow and result of the identification of the origin of a
primary cancer. The cluster analysis identified the lymph node metastasis
microRNA profile as colorectal cancer. All the colorectal samples, including
the metastasis, cluster together and clearly indicate that the colon was
the origin of the metastasis.
21
22
LIQUID HANDLING TECAN JOURNAL 1/2009
Automation accelerates
crop improvement
Scientists at the Institute of Plant Breeding of the University
of Kiel, Germany, are developing TILLING® (Targeting Induced
Local Lesions in Genomes) programs in rape seed and sugar
beet, relying on their Freedom EVO® liquid handling workstation
for high throughput DNA purification and normalization using
MACHEREY-NAGEL NucleoSpin® 96 Plant DNA extraction kits.
The Institute of Plant Breeding, part of the
Institute of Agronomy and Crop Science
at the University of Kiel, is a research and
teaching facility, conducting various plant
breeding projects in crop species like sugar
beet, barley, rape seeds and asparagus.
Some of the projects are funded by the
German national genome program, Genome
Analysis of the Plant Biological System
(GABI), including rape seed and sugar beet
TILLING projects. Tecan’s Freedom EVO
200 liquid handling workstation is being
used for automated DNA extraction and
normalization for establishing the TILLING
projects in which, typically, there are 4,000
or more plants in a population from which
DNA needs to be individually extracted.
Prof Christian Jung and
Mrs Gislind Bräcker
“We chose Tecan’s workstation because we
needed an automated, high throughput
system to extract DNA,” explained Professor
Christian Jung, Director of the Institute of
Plant Breeding.
“Our Freedom EVO platform has an
integrated centrifuge, so it is able to perform
DNA purification using MACHEREY-NAGEL’s
NucleoSpin 96 Plant kit, automating the
whole procedure after homogenization
of leaf material with a SPEX SamplePrep
2000 Geno/Grinder®, which uses small
steel beads to process simultaneously up
to 48 samples in 2 ml reaction tubes, or
192 samples using 8-strip microtubes in
a 96-well format. The Freedom EVO also
normalizes the purified DNA, using the
integrated GENios™ fluorescence reader
to measure the DNA concentration in
96-well microplates and dilute the DNA
in each well to the right concentration for
PCR amplification. This is a huge help for
our projects, because it is very tedious to
normalize by hand, and almost impossible in
micro plates, so the automation makes life
much easier for the technician faced with
normalizing thousands of DNA samples. The
DNA needs to be of good quality, because
it will be stored and used in PCR reactions
for the next 10 to 20 years. DNA extracted
manually may not be of sufficient quality
to withstand long-term storage whereas, in
combination with the MACHEREY-NAGEL
extraction kits, we can obtain highly purified
DNA that we can store for long periods. The
greatest advantage of this system is that
the entire purification and normalization
procedures are automated, and this offers
the technician time to perform other tasks.”
The institute has had the Freedom EVO since
2006, and Tecan and MACHEREY-NAGEL
worked together to adapt and optimize the
protocol for automated DNA extraction and
normalization. Prof Jung added: “We have
been using it routinely for about a year now,
and our technician, Gislind Bräcker, is happy
with its performance. The yield of DNA has
LIQUID HANDLING TECAN JOURNAL 1/2009
(l) The Institute of Plant Breeding
at the University of Kiel
(r) The Freedom EVO platform is
used for high throughput DNA
purification and normalization
been improved, and we can consistently
purify several micrograms of DNA from each
sample, processing up to 192 samples in 2 ml
reaction tubes in a day. The complete DNA
extraction process takes about 6.5 hours,
including the four hours spent to prepare
the starting material by homogenization of
leaf tissue, removal of the steel beads, and
a two-hour incubation and centrifugation.
In the future, we are planning to switch to
using 8-strip microtubes in 96-well format,
with which we expect the procedure to
be quicker. The PCR set-up will also be
automated on the Freedom EVO in the
future, to further streamline our workflow.
For DNA normalization, 384 samples can be
measured and adjusted in one day.”
TILLING involves the identification of
mutants within specific genomic regions,
among huge populations that have
been mutagenized by ethylmethane
sulfonate (EMS). M1 generation plants
are mutagenized and self-pollinated to
produce the M2 population, and the DNA
from the leaves of each individual M2
plant is extracted and stored, while the
plants are self-pollinated again and M3
generation seeds are produced. The DNA is
analyzed by PCR in specific genomic regions
of interest, and the amplified fragments
are treated with endonuclease CelI, which
cleaves mis-matches, yielding additional
fragments when there is a base substitution
in the selected region. When the mutated
sequence indicates the possibility of an
altered phenotype, the corresponding
batch of M3 seeds is analyzed, and the
plants carrying the mutation are isolated
for further study. “Plant breeders need
new genetic variation and, because of the
unpopularity of transgenic technology in
Europe, mutation technology and TILLING is
a new source for genetic variation. Breeders,
particularly in Europe, are very interested
in this technology. Identifying sequence
variants or mutations in this way, by
genotype rather than by phenotype, is now
possible because more and more sequence
information of crop species is available.
That is the reason why a number of these
TILLING platforms have been established
in different countries around the world for
various crop species.”
Prof Jung continued: “We are currently
developing TILLING platforms for rape
seed and sugar beet, in collaboration
with commercial plant breeders. For one
project in particular, we are looking at a
deleterious component in rape seed, called
sinapin, that makes it unsuitable as animal
fodder. We are looking for plants that have
a mutation in one of the genes for sinapin
metabolism, and we expect that if one of
these major genes is mutated, the plant will
have a reduced amount of sinapin, and this
mutant would then be suitable for feeding
to animals. This is a clear practical breeding
purpose, where the yield of our project can
be used by commercial plant breeders.”
“Very recently, the animal breeding unit
of the university has started to use the
Freedom EVO for automated DNA extraction
from cows for a program on mapping,
genotyping and haplotyping cattle DNA.
Although between us we have huge
numbers of DNA extractions to perform,
the Freedom EVO has the capacity to cope
with these sample numbers,” concluded
Prof Jung.
Geno/Grinder is a registered trademark of SPEX
SamplePrep, Inc.
NucleoSpin is a registered trademark of MACHEREYNAGEL, GmbH & Co.
TILLING is a trademark of Arcadia Biosciences, Inc.
23
24
DETECTION TECAN JOURNAL 1/2009
Getting under the skin
of stem cells
Researchers in the Center for
Biosciences at the Karolinska
Institute in Stockholm,
Sweden, have been using an
Infinite® 200 microplate
reader and NanoQuant Plate™
for the characterization of a
newly isolated population of
skin stem cells in mice.
Researchers in the Center for Biosciences
at the Karolinska Institute, Sweden, have
recently isolated skin stem cells displaying
several exciting properties which challenge
previous assumptions. The study, published
in Nature Genetics*, demonstrated that
these stem cells (Lgr5+ cells) are the most
primitive skin stem cells yet described, able
to divide actively and migrate through
the skin tissue during follicle growth and
regression. These cells are partly regulated by
the well characterized ‘Hedgehog’ signaling
pathway, and are of particular interest to the
wound-healing process and the development
of basal cell carcinomas (BCC), the most
prevalent form of skin cancer.
Characterization of gene expression in
these cells has been particularly difficult
due to their scarcity. Dr Maria Kasper, from
the Karolinska Institute’s environmental
toxicology group, explained: “These cells
are extremely rare, and so the experimental
material we obtain is especially valuable;
100,000 sorted cells only renders 150 ng of
RNA. Once this material has been isolated,
it is used to identify signaling pathways
involved in stem cell regulation by qPCR.
The significance and reliability of the data
obtained is dependent on both the quantity
and integrity of the extracted RNA, as qPCR
using very low starting concentrations of RNA
is particularly sensitive to these variables.
Therefore, it is very important for us to
achieve exact quantification of extracted
samples with minimal loss of material.”
Traditional photometry-based systems for
evaluating RNA samples are dependent on
comparing absorbance measurements at
multiple wavelengths, but most readers
cannot produce reliable absorbance
readings from small volumes of RNA sample.
However, using Tecan’s NanoQuant Plate in
conjunction with a multi-functional Infinite
200 microplate reader offers the necessary
sensitivity for reliable low volume RNA
purity analysis. Maria commented: “Using
this system ensures that only minimal
material is required for reliable evaluation of
our valuable RNA samples and allows us to
investigate more than 30 selected genes by
qPCR from the initial 150 ng of isolated RNA.”
Lgr5 expression in the mouse hair follicle.
Immunohistochemical staining showing
localization of Lgr5 (blue) and CD34 (brown)
expressing cells.
DETECTION TECAN JOURNAL 1/2009
The NanoQuant Plate is a 16-channel
quartz optic for measurement of low
concentrations and low sample volumes,
with quick and simple washing procedures
to eliminate cross-contamination.
Specifically designed to give outstanding
performance and a very high rate of
reproducibility, the NanoQuant Plate is
intended for a broad range of applications
requiring sample volumes as low as 2 µl.
Combined with the Infinite 200 microplate
reader, this system allows accurate
absorbance measurement of DNA/RNA
concentrations as low as 1ng/μl, helping to
conserve valuable samples.
Maria Kasper, Takashi Shimokawa and Viljar Jaks
Maria’s colleague, Dr Viljar Jaks, continued:
“Having the NanoQuant Plate in the
laboratory has been very useful for our
investigations. Because it is integrated into
the Infinite microplate reader, it is very
easy to use and the i-control™ software
is intuitive. Although our studies are not
high throughput by nature, the 16-channel
plate format allows us to analyze entire
experimental sets together.”
“Accurate quantitation allows us to
normalize RNA concentrations very precisely
for further analysis of gene expression using
a variety of methods, helping to identify
the pathways contributing to the behavior
of these stem cells. Elucidation of these
regulatory mechanisms could help us to
understand the role of keratinocyte stem
cells in various forms of tissue growth.
One example of this is the wound healing
process. Because hair follicles are laid down
during embryonic development, they are not
formed in human scar tissue when new skin
is grown. Therefore, the next important step
is to identify a similar stem cell population
in human hair follicles. If we can understand
how these stem cell populations are
regulated, then we may be able to induce
proliferation to produce new hair follicles.”
Maria added: “Another application of great
interest is the development of basal cell
carcinoma – the most common form of
skin cancer – the origins of which have not
yet been determined. It is still not clear
whether BCC originates in the interfolicular
epidermis or the hair follicles, but using Lgr5
as a marker of hair follicle stem cells offers a
potentially powerful tool to investigate the
genesis of this cancer. Deregulation of the
pathways that control these cells can cause
sustained activation of their proliferation
and disturb their normal differentiation
process, which increases the risk of cancer
formation.”
(* Nature Genetics 40, 1291 - 1299 (2008))
The Infinite 200 and NanoQuant Plate for
analizing small volumes of RNA sample
25
26
SAMPLE MANAGEMENT TECAN JOURNAL 1/2009
Automated processing
for pioneering translational
medicine project
Aurora Health Care is relying
on its Freedom EVO® liquid
handling platforms and REMP
Small Size Store™ to provide
automated processing,
cataloging and storage of
samples for an innovative
translational medicine project.
Aurora Health Care is a not-for-profit health
care provider, serving eastern Wisconsin in
the United States. Established in 1984 by
the affiliation of two Milwaukee hospitals,
Aurora Health Care was founded on a simple
premise; that there is a better way to provide
health care. Since these early days, Aurora
has expanded to serve a population of more
than a million people, and now comprises
13 hospitals, over 120 outpatient clinics and
approximately 140 pharmacies.
Aurora’s founding principle still remains
central to its vision for health care, and has
been the driving force behind establishing
a translational medicine project – a
fully automated repository of biological
information developed over the last five
years by Aurora directors Matthew and
Alfred Tector. With so many facilities, in
addition to community services, including
visiting nurses, physicians and hospice
activities, the potential influence of
this venture is far reaching. The Aurora
biorepository hopes to collect 50,000
samples a year, making use of surplus blood
samples collected from patients for routine
clinical tests that would otherwise go to
waste. With such a large volume of samples
for processing, automated sample handling
is essential to its success.
Dr Matt Tector, explained: “The aim of the
project is to allow this research to take place
without changing the experience of patients
within the health care system. We will be
asking patients entering the system, as part
of the preliminary paperwork, to consent
to letting us access their medical records
and use surplus blood samples. The benefit
of this arrangement is that patients won’t
have to give additional blood samples, so
we expect a high consent rate. Potentially
we could create a library on the same scale
as the UK Biobank, benefiting all races and
socio-economic backgrounds within our
region. To process such a large number of
samples, we will need to use robotics for as
much of the processing as possible. All our
colleagues with experience of automated
systems recommended Tecan as the right
choice for liquid handling. With the Freedom
EVO/REMP SSS Factory, almost the entire
process will be automated.”
“The objective of establishing the Aurora
biorepository is not the discovery of new
drugs and therapies, but the improvement
of existing treatments,“ Matt continued.
“As a health care system, our advantage
is that we are not limited to investigating
a single disease, like some institutions.
Our aim is to transfer information from a
research environment to a clinical setting as
fast as possible. We intend to have people
in the front line of health care – clinicians,
nurses, physicians and pharmacists –
directing our research to the appropriate
studies. We can then find the expertise we
need to help us accomplish those goals.
If we make a discovery that is relevant to
any aspect of the health care industry, our
unique set-up allows us to implement those
changes rapidly.”
“Our intention is to use genetic analysis to
help our physicians to create personalized
therapies for individual patients. We are not
a clinical laboratory, however, so we will not
be reporting results to directly influence
an individual donor’s care. In fact, one of
the tasks of the Freedom EVO will be to
assign new identities to each sample so
they cannot be traced back to identify the
patient. We have a Freedom EVO 100 that
will be responsible for checking the barcodes
on samples against the database. It will then
re-rack those samples from patients who
have consented to this project, and assign
a new identity which incorporates certain
elements of their medical history. With this
system we expect to be able to process
several thousand samples a day. Samples
will then be transferred to our Freedom EVO
SAMPLE MANAGEMENT TECAN JOURNAL 1/2009
(l to r) Matt Kirsling, information systems specialist, Alfred Tector, MD, clinical research and transplant program medical
director, Natalie Polinske, research associate and Matt Tector, PhD, biorepository director
200 / REMP SSS Factory, which will isolate
DNA from the samples and store them for
later use. Full automation of the process
should allow us to process approximately
300 DNA samples a day. We have also
purchased a Safire2™ microplate reader and
are hoping to incorporate some standard
genetic analysis assays into the Freedom
EVO 200 protocols.”
“Planning for the biorepository has been
ongoing for five years, and our relationship
with REMP began very close to the start
of the process. As the project has evolved,
the Tecan team has always offered good
insight; they have put almost as much into
this project as we have. Whenever we have
needed help, Tecan’s support staff has been
there. As the project grows and expands to
our other facilities we hope to continue our
relationship with Tecan.”
27
28
LIQUID HANDLING TECAN JOURNAL 1/2009
Signaling success
The Center for High
Throughput Cell Biology,
a brand new facility at Yale
University, Connecticut, is
combining automation with
siRNA screening techniques
to investigate signal
transduction pathways.
With the aim of ‘re-writing
the book’ the laboratory aims
to use the high throughput
capacity of its Freedom EVO®
workstations to conduct
genome-wide screens of
primarily HUVEC cells to
validate new and existing
data.
Professor James E Rothman, recently
returning to his alma mater as Chair of the
Department of Cell biology at Yale University,
has been instrumental in establishing the
Center for High Throughput Cell Biology,
a state-of-the-art facility situated at Yale’s
new research campus between West Haven
and Orange, Connecticut. Professor Rothman
is no stranger to high throughput screening
programs, having previously been involved
in the NIH Roadmap initiative as Director of
the Genome Center of Columbia University
in Manhattan, New York. The Center for High
Throughput Cell Biology is a facility affiliated
to the Department of Cell Biology, under the
directorship of Dr Lars Branden – a former
associate director of the Genome Center at
Columbia – and is supporting its ambitious
research program with provision of
screening services for external laboratories.
Lars explained the aims of the Center’s
research: “The basic question we are trying
to answer is ‘How are signal transduction
networks organized?’ The intention is to
identify the signal transduction network
connections between the different signaling
pathways, and the components within
each pathway. Our approach differs from
other projects in that we are primarily
looking at one specific cell type, human
umbilical vein endothelial cells (HUVEC)
for our internal research. This is because
many of these transduction pathways have
been elucidated in universal terms, but
the theoretical pathway often does not
agree with experimental findings when
you get down to specific cell types. We are
using pooled HUVEC samples to ensure
specific genetic variations do not skew our
findings, and are hoping to take in all the
information about signal transduction from
existing databases and validate it for our
experimental set-up.”
“The primary technology we are using to
identify the key regulatory entities within
different signaling pathways is genomewide screens with siRNAs. If we identify
a particular gene we want to know more
about, we can run it against all the signal
transduction pathways we have established
assays for. We have already developed
over 150 high-content assays in-house and
about 50 low-content assays. If we find a
gene that perturbs a specific pathway, we
then perform extensive secondary assays,
‘biological profiling’, to identify the more
specific biological relevance of the gene in
question. Combining these techniques to
identify the key regulators of each pathway
is our initial goal, however, this is really a
stepping stone for trying to understand the
connections between pathways. From there
we can look at how this ‘transcriptional
network’ varies between cell types. If you can
understand these differences then this could
help to explain why you get side effects from
LIQUID HANDLING TECAN JOURNAL 1/2009
a certain drug, or how paracrine signaling
works in organogenesis.”
and this cannot be achieved if the process
requires too much human intervention.”
Lars added: “A project of this scale requires
a multi-disciplinary approach, so within the
Center we have units responsible for assay
development, high throughput screening
and informatics. It is hard to separate the
biological science from the technology
for this project, as the two are intrinsically
linked. The project would be impossible
without automation, and Tecan really
cares about what we are doing and listens
to our needs. We brought one Freedom
EVO platform with us from Columbia
University, and this unit is dedicated to
384-well plate formats. Tecan is currently
building three new platforms for us, two
assay workstations and an immunohistochemistry workstation, which will
all be running 1,536-well plates. These
systems will greatly improve our throughput
compared with our previous set-up. When
first designing our system four years ago,
we integrated many functions into a single
platform, and we have continued down that
route with our new units. The Freedom EVO
platforms are very flexible, offer amazing
precision and, by using a modular approach,
we can optimize the space available on
the deck of the workstation. The plates are
all in racks to simplify transfer between
workstations; the whole point of automated
integration is that it gives you total control
“The new 1,536-format platforms should be
able to run up to 80 plates in 24 hours and,
if you consider that a genome-wide screen
in quadruplicate only requires about 52
plates, this really demonstrates our potential
capacity. Our new imaging workstation also
has facilities to conduct both primary and
secondary screening assays so, if we get a
positive result, we can re-screen the well at
high resolution and run secondary assays
we have built into the system. This multiplex
approach helps to minimize cost and
increase efficiency, generating a lot of data
very rapidly, and our biggest bottleneck will
then most definitely be in data storage and
data processing.”
“Working closely with Tecan to design this
system, we have realized that we would
like to take our collaboration to the next
level. Tecan has believed in this work from
the beginning and, even before a proposal
was put to us, there was a lot of joint work
done to resolve potential problems. It has
to be a two-way relationship and, from the
very beginning, we have been developing
some new ideas together. It’s a lot easier
to work with a company with whom you
have established a good rapport and, with
Tecan, we know that we will always get the
support we need.”
29
30
DETECTION TECAN JOURNAL 1/2009
Collaboration paves
the way for flexibility in
diagnostic laboratories
Applications specialists from Tecan Austria have teamed up
with scientists from Technoclone GmbH, Vienna and the
University Children’s Hospital in Zurich to set up fast and
flexible microplate-based assays for a full range of benign
hematology conditions in children.
The University Children’s Hospital in
Zurich is Switzerland’s largest pediatric
facility, and offers a spectrum of pediatric
specialties ranging from emergency care
through to surgery, as well as boasting an
extensive research program. The hospital’s
hematology division combines routine
clinical diagnostics with investigation into
a range of erythrocyte- and platelet-related
conditions in children. Dr Oliver Speer,
head of the Experimental Hematology and
Erythrocyte Laboratory, explained: “Our
work is focused on benign hematology.
This includes hemaglobinopathies such
as thalassemia and sickle cell disease, as
well as enzyme defects in the erythrocyte
glycolytic pathway, such as G6PD deficiency
(Favism) or ADA (important for diagnosis of
certain immune deficiencies). We also look
into primary and secondary hemostasis,
including a major research project
examining the endogenous thrombin
potential before and after therapy in
paediatric idiopathic thrombocytopenia
purpura (ITP) patients. We work exclusively
with patient samples, performing analyses
with whole blood, platelet-rich plasma,
plasma or cell culture depending on
the investigation.”
Many of the laboratory’s assays, for
both routine diagnostics and research
applications, are based on photometric
techniques and historically the laboratory
used different instruments for each assay.
In the last year, Dr Speer has overseen
the transfer of all photometry to Tecan’s
Infinite® M200 Quad 4 Monochromators™based microplate reader: “This new system
is faster than our previous set-up and we
can use it for all of our assays. We have
gone from using 1 cm cuvettes to 96-well
microplates and this gives us so much more
flexibility, as well as the speed and capacity
for a much higher sample throughput. The
protocol for each assay is pre-set, so all we
need to do is select the relevant protocol
and the system automatically gives onscreen instructions. Initially we compared
results between the Infinite and our
older instruments and we were very soon
confident enough to switch entirely to the
Tecan instrument. Once we had completed
transfer and validation of our existing
assays, we realised that the flexibility and
sensitivity of the Infinite reader would allow
us to supersede some of these assays with
more efficient or sensitive methods. One
example of this is ferritin detection,
(front l to r) Marlis Schmid, Karin Zurbriggen,
(back l to r) Oliver Speer, Alexandra Förderer, team
members of the hematology laboratory
which was previously assessed using an
ELISA technique. With the new system
we have been able to switch from this
expensive and time-consuming method to
a far faster and more flexible turbidometric
assay. We also became interested in
new technologies such as Technoclone’s
thrombin generation assay (TGA).”
Technoclone’s novel TECHNOTHROMBIN®
TGA measures thrombin formation
using a fluorigenic substrate and can be
used to monitor hemophiliacs during
inhibitor-bypassing therapy, to monitor
anticoagulation therapy and to determine
states of thrombophilia. Veronika Binder,
CEO of Technoclone, explained: “The
TECHNOTHROMBIN TGA offers exceptional
sensitivity. It measures the whole kinetics
of thrombin generation, not only during the
initial phase of thrombin formation, but
also during the phase of down-regulation
and inactivation of the formed thrombin.
Dr Speer approached us requesting
validation data for the assay using the
Infinite system and, because so many of
our customers have Tecan instrumentation,
we recognized that validating it for the
Infinite series readers would be of great
value to many laboratories. It also offered
us a rare opportunity to work with pediatric
samples, so we approached Tecan and
worked closely together on the validation.
Application specialists from Tecan Austria
validated the instrument using control
samples and soon Dr Speer was able to
continue the process using patient material.
Tecan has been very good to work with, and
this has been an excellent example of two
companies working together for the benefit
of a joint customer. Validating our assay
with Tecan instrumentation really does
provide customers worldwide with the best
diagnostic solution.”
DETECTION TECAN JOURNAL 1/2009
Dr Speer added: “The collaboration between
the companies has been perfect for us. Staff
from Tecan and Technoclone came to the
hospital, and we were able to set things up
very easily; I have been surprised at just how
quickly we have managed to complete this
study. Throughout the entire process, Tecan
has provided excellent support, from the
initial installation of our system to further
developing the software specifically for our
needs. We are regularly in contact regarding
technical questions and I very much enjoy
working with the Tecan team.”
TECHNOTHROMBIN is a registered trademark of
Technoclone GMBH.
Kinetics of thrombin generation
Leading the debate
‘Leading the debate’ is a new feature to the Tecan
Journal where we invite you to share your views about
trends in life sciences and the newest hot topics for
our industry. Marc Feiglin, Chief Technology Officer
for Life Sciences, gives us a flavor of what’s to come in
forthcoming issues.
“Here at Tecan we are in an enviable position to be working
closely with many leading luminaries in the life sciences world.
Most recently, in September 2008, we hosted the second
annual Tecan symposium, bringing together researchers
from around the world who rarely get the opportunity in
their busy schedules to thrash out the issues affecting their
own particular area of science. The focus of this event was
biotherapeutics, stem cells, and regenerative medicine, rapidly
developing areas that are grabbing the attention of life science
researchers and clinicians alike. There are many outstanding
questions associated with these technologies. How will
the regulatory aspects for biosimilars develop compared
to those that currently exist for small molecule-based
therapeutics? Have we begun to see an impact yet from stem
cell-based therapies? Where are the market opportunities for
regenerative medicine? Will the recent political change in the
US and the possibility of a major investment program there
open up these areas even further? What role can companies
like Tecan play in developing these opportunities further?”
“We certainly had some lively conversations on
these and related topics during the three-day
symposium in September and would love to hear
your opinions too! If you’d like to contribute to our
‘Leading the debate’ column, send us your thoughts
by email to talk@tecan.com.”
The thrombin generation cascade
31
32
EVENTS 2009 TECAN JOURNAL 1/2009
Meet Tecan at these events in the first part of this year
Americas
Lab Automation 2009
Palm Springs, CA
25-27 Jan 2009
ABRF 2009
Memphis, TN
07-10 Feb 2009
MD&M West
Anaheim, CA
09- 12 Feb 2009
AAFS 61st Annual Scientific Meeting
Denver, CO
16-21 Feb 2009
Molecular Medicine Tri-Conference
San Francisco, CA
25-27 Feb 2009
60th Pittsburgh Conference on Analytical Chemistry
And Applied Spectroscopy (Pittcon)
Chicago, IL
08-13 Mar 2009
AACR 100th Annual Meeting
Denver, CO
18-22 Apr 2009
34th Lorne Conference on Protein Structure and Function
Lorne, Australia
08-12 Feb 2009
Japanese Society for Bioscience, Biotechnology and Agrochemistry
Fukuoka, Japan
28-29 Mar 2009
Asia and Pacific
Europe, Middle East and Africa
CHUV Research Day 2009
Lausanne. Switzerland
29 Jan 2009
MNBS Contact Meeting 2009
Røros, Norway
29 Jan 2009
LogP 2009
Zurich, Switzerland
08-11 Feb 2009
29. Spurenworkshop der Deutschen Gesellschaft für Rechtsmedizin
Munster, Germany
27 Feb 2009
Statusseminar Chip Technologies 2009
Frankfurt, Germany
5-6 Mar 2009
Laborama
Brussels, Belgium
12 Mar 2009
BioProcess
Düsseldorf, Germany
22-23 Apr 2009
AAC Spring Meeting 2009
Edinburgh, UK
27-28 Apr 2009
Tecan. For all your lab automation needs.
Liquid Handling & Robotics | Detection | Sample Management | Components | Services & Consumables
Headquarters:
Tecan Group Ltd., Seestrasse 103, CH-8708 Männedorf, Switzerland
T +41 44 922 88 88 F +41 44 922 88 89 info@tecan.com
Tecan
Austria +43 62 46 89 33 Belgium +32 15 42 13 19 China +86 10 586 95 936 France +33 4 72 76 04 80 Germany +49 79 51 94 170 Italy +39 02 92 44 790
Japan +81 44 556 73 11 Netherlands +31 18 34 48 17 4 Portugal +351 21 000 82 16 Singapore +65 644 41 886 Spain +34 93 490 01 74
Sweden +46 31 75 44 000 Switzerland +41 44 922 89 22 UK +44 118 9300 300 USA +1 919 361 5200 ROW +43 62 46 89 33
REMP
Switzerland +41 31 770 70 70 Japan +81 44 542 70 21 USA +1 508 429 2200
Tecan Journal, Customer Magazine of Tecan Trading AG., ISSN 1660-5276
Design: OTM/London www.otmcreate.com
Photography: Marc Wetli/Zürich www.wetli.com, Günter Bolzern/Zürich www.bolzern.net, Susanne
Völlm/Zürich www.susannevoellm.ch
Editor in Chief: Tecan Trading AG, Cornelia Kegele
Project Lead: Tecan Trading AG, Cornelia Kegele
Editor: kdm/UK www.kdm-communications.com
Print: DAZ Druckerei Albisrieden AG/Zurich www.daz.ch
Address: Tecan Trading AG, Marketing Communications, Seestrasse 103, CH-8708 Männedorf,
Switzerland, journal@tecan.com,www.tecan.com
Tecan Group Ltd. makes every effort to include accurate and up-to-date information within this
publication, however, it is possible that omissions or errors might have occurred. Tecan Group Ltd.
cannot, therefore, make any representations or warranties, expressed or implied, as to the accuracy
or completeness of the information provided in this publication. Changes in this publication can be
made at any time without notice. All mentioned trademarks are protected by law.
www.tecan.com
For technical details and detailed procedures of the specifications provided in this document
please contact your Tecan representative.
This brochure may contain reference to applications and products which are not available in all
markets. Please check with your local sales representative.
8 Plus 1 Access, ABS, Gemini, Genesis RSP, GENios, HS 400 Pro, HS 4800 Pro, i-control, Instant
Pipetting, MCA, MultiChannel Arm, NanoQuant Plate, OSND, PosID, Power Washer 384, Quad4
Monochromators, Safire2, Te-MO and Te-Stack are trademarks and Cavro, Freedom EVO, Freedom
EVOware and Infinite are registered trademarks of Tecan Group Ltd, Männedorf, Switzerland.
Tecan is in major countries a registered trademark of Tecan Group Ltd., Männedorf, Switzerland.
REMP Small-Size Store is a trademark of REMP AG, Oberdiessbach, Switzerland.
© 2009 Tecan Trading AG, Switzerland, all rights reserved.